Chrysosporium cellulase and methods of use

Abstract

A computer-based method and apparatus for the analysis specification and support of work processes. The system is designed to support multiple interdependent decisions, at least some of which require collaboration among multiple participants ( 116 ). Work processes are modeled using an application framework ( 99 ) used to develop abstract, decision ( 100 ) process models. The decision ( 100 ) process models are used as a pattern to instantiate concrete process models that incorporate the work defined by the abstract process. The process model is then used to instantiate project models that incorporate the required work from the process. The project models are used to direct and guide the behavior of the participants ( 116 ) in the work process.

1. An isolated culture of  Chrysosporium lucknowense  Garg 27K having accession number VKM F-3500D. 
     
     
       2. A composition comprising an isolated neutral and/or alkaline cellulase, said isolated neutral and/or alkaline cellulase is obtained from a wild type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D. 
     
     
       3. A composition according to  claim 2 , wherein the fungus is a mutant strain of  Chrysosporium lucknowense  Garg 27K. 
     
     
       4. A composition according to  claim 2  having cellulase activity at a temperature from about 40° C. to about 60° C., at a pH from about 5.0 to about 11.0. 
     
     
       5. A composition according to  claim 2  having at least 50% of the optimal cellulase activity, at a pH from about 6.0 to about 7.0, at a temperature from about 40° C. to about 60° C. 
     
     
       6. A composition according to  claim 2  wherein said cellulase activity is assayed by any one of the CMCase, RBBCMCase, endoviscometric or filter paper activity assays. 
     
     
       7. A substantially purified and isolated protein fraction, obtained from a composition according to  claim 2 , and having at least 50% of its maximal cellulase activity at a pH between about 6.0 and about 7.0. 
     
     
       8. An endoglucanase obtained from a fraction according to  claim 7 , having a molecular weight of about 25 kD and pI of about 4. 
     
     
       9. An endoglucanase obtained from a fraction according to  claim 7 , having a molecular weight of about 70 kD and a pI of about 4. 
     
     
       10. An endoglucanase obtained from a fraction according to  claim 7 , having a molecular weight of about 60 kD and a pI of about 3. 
     
     
       11. An endoglucanase obtained from a fraction according to  claim 7 , having a molecular weight of about 43 kD and a pI of about 3. 
     
     
       12. A cellobiohydrolase obtained from a fraction according to  claim 7 , having a molecular weight of about 60 kD and a pI of about 4. 
     
     
       13. A substantially purified and isolated neutral and/or alkaline cellulase enzyme, isolated from a protein fraction according to  claim 7 , and having a pI of between about 3 and about 5.5. 
     
     
       14. A cellulase according to  claim 13  wherein said cellulase possesses either endoglucanase or cellobiohydrolase activity. 
     
     
       15. A cellulase according to  claim 13  wherein said cellulase retains at least 50% of its maximal cellulase activity at a pH between about 6.0 and about 7.0. 
     
     
       16. An endoglucanase obtained from a fraction according to  claim 7  and having a molecular weight of about 25 kD. 
     
     
       17. An endoglucanase obtained from a fraction according to  claim 7  and having a molecular weight of about 70 kD. 
     
     
       18. An endoglucanase obtained from a fraction according to  claim 7  and having a molecular weight of about 43 kD. 
     
     
       19. A detergent composition containing one or more purified enzymes isolated from a protein fraction according to  claim 7 , and further comprising a surfactant. 
     
     
       20. A fabric softening composition containing one or more purified enzymes obtained from the protein fraction according to  claim 7 . 
     
     
       21. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, comprising an isolated cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D. 
     
     
       22. A composition according to  claim 21  further comprising one or more components selected from the group consisting of pumice stones, abrasives, softeners, solvents, preservatives, bleaching agents, bluing agents, fluorescent dyes, antioxidants, solubilizers, detergents, surfactants, enzymes, builders, anti-redeposition agents, buffers, caking inhibitors, masking agents for factors inhibiting the cellulase activity, and cellulase activators. 
     
     
       23. The composition of  claim 21 , wherein the pH is between 10.0 and about 11.0. 
     
     
       24. The composition of  claim 22 , wherein the pH is between 10.0 and about 11.0. 
     
     
       25. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, comprising an isolated cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D, said composition further comprising one or more components selected from the group consisting of proteinases, detergents, and surfactants. 
     
     
       26. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least about 33 units of endo-1,4-β-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of an isolated cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D. 
     
     
       27. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least about 33 units of endo-1,4-β-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of an isolated cellulase obtained from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D. 
     
     
       28. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least about 120 units of 1,4-β-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of an isolated cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D. 
     
     
       29. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least about 120 units of endo-1,4-β-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of an isolated cellulase obtained from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D. 
     
     
       30. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least about 964 units of endo 1,4-β-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of an isolated cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D. 
     
     
       31. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least about 964 units of endo-1,4-β-glucanase activity per gram of dry composition, of an isolated cellulase obtained from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K. accession number VKM F-3500D. 
     
     
       32. A laundry detergent composition, comprising an isolated cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus  Chrysosporium  wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D, further comprising one or more surfactants. 
     
     
       33. A laundry detergent composition, comprising an isolated cellulase obtained from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K accession number VKM F-3500D, further comprising one or more surfactants. 
     
     
       34. A method for producing the composition according to  claim 2 , said method comprising growing a mutant fungus of the genus  Chrysosporium  in culture in a suitable medium, wherein the neutral and/or alkaline cellulase is secreted into the medium. 
     
     
       35. A method of stonewashing denim fabric or denim jeans, said method comprising treating said denim fabric or denim jeans with a composition according to anyone of  claims 3 ,  7 ,  21 , and  2 , thereby obtaining a stonewashed denim fabric or denim jeans. 
     
     
       36. A method of biopolishing, defibrillating, bleaching, dyeing, or desizing textiles comprising treating said textiles with a composition according to anyone of  claims 3 ,  7 ,  21 , and  2 , thereby obtaining a textile which has been polished, bleached, dyed, desized or has reduced fibers. 
     
     
       37. A method of deinking or biobleaching paper or pulp, said method comprising treating said paper or pulp with a composition according to anyone of  claims 3 ,  7 ,  21 , and  2 , thereby obtaining a paper or pulp which has been deinked or bleached. 
     
     
       38. A method for enhancing the softness or feel of cellulose or cotton-containing fabric, comprising treating said fabric with a composition according to anyone of  claims 3 ,  7 ,  21 , and  2 , thereby obtaining a softer fabric. 
     
     
       39. A method for generating mutant strains of the genus  Chrysosporium  which produce enhanced cellulase activity at neutral and/or alkaline pH's, comprising:
 (a) mutating spores of a fungus of the genus  Chrysosporium;    
 (b) culturing the spores from step (a); and 
 (c) screening the cultures from step (b) for enhanced levels of neutral and/or alkaline cellulase activity, wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D, said cultures have cellulase activity at a temperature between 40° C. and 60° C., and at a pH of about 5 to about 12. 
 
     
     
       40. The method of  claim 39  wherein the mutating step comprises exposing the spores to ultraviolet light or a chemical mutagen. 
     
     
       41. The method of  claim 40  wherein the chemical mutagen is nitrous acid, N-methyl-N′-nitro-N-nitrosoguanidine, or 4-nitroquinolone-N-oxide. 
     
     
       42. A mutant strain of the genus  Chrysosporium  obtained by the method of anyone of  claims 39  or  40 . 
     
     
       43. A method of culturing a fungus of  Chrysosporium lucknowense  Garg 27K having accession number VKM F-3500D in a medium containing inorganic salts, carbon sources, and organic nitrogen sources, at a pH between about 5 and 8. 
     
     
       44. A method of culturing a fungus of the genus  Chrysosporium  according to  claim 43 , wherein the pH is between about 6.5 and 7.5. 
     
     
       45. A method of culturing a fungus of the genus  Chrysosporium  according to  claim 43 , wherein the pH is between about 6.9 and 7.1. 
     
     
       46. A method of culturing a fungus of the genus  Chrysosporium  according to  claim 43 , wherein the pH is maintained at 7.5. 
     
     
       47. The composition according to  claim 3 , wherein the fungus is a mutant strain of  Chrysosporium lucknowense  Garg 27K having accession number VKM F-3632D. 
     
     
       48. A method of saccharification of cellulose, comprising treating the cellulose with an isolated cellulase enzyme of  claim 13  having at least 50% of its maximal cellulase activity at a neutral and/or alkaline pH, whereby the cellulose is hydrolyzed. 
     
     
       49. The method of  claim 48 , wherein the cellulose is a lignocellose biomass from agriculture, forest products, municipal solid waste, or other like sources. 
     
     
       50. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least about 191 units of endo-1,4-β-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of an isolated cellulase whose amino acid sequence is encoded by a nucleic acid sequence from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D. 
     
     
       51. A composition for the enzymatic treatment of cellulosic fibers or cellulosic fabrics, having at least about 191 units of endo-1,4-I3-glucanase activity per gram of dry composition, as measured by an endoviscometric assay, of an isolated cellulase obtained from a wild-type or mutant fungus of the genus  Chrysosporium , wherein the fungus is  Chrysosporium lucknowense  Garg 27K, accession number VKM F-3500D.